Extension of shelflife of the fermented fish product, shidal by packaging in glass bottle and low temperature storage

Abstract

Shidal is a very popular, salt free, semi-fermented traditional fish product of north-east India prepared using minor carps
(Puntius spp.) in specially designed earthen pots (mutka). The prime quality of shidal is given by its characteristic aroma and
flavor which is lost very fast, once it is taken out of mutka. A study was conducted to preserve the quality of shidal outside
mutka, by packing in glass bottles and storing under refrigerated temperature. Microbiological, biochemical and sensory changes
during storage period of 120 days were analysed at 15 days interval. The total plate count (TPC) did not change significantly
(p>0.05) and remained near 7 log CFU g-1 during storage. The total fungal count (TFC) was negligible., The pH of shidal was
initially acidic (4.42 ± 0.25) which increased significantly (p<0.05) towards the end of storage period. The non-protein nitrogen
(NPN), free α-amino nitrogen (AAN) and total volatile base nitrogen (TVB-N) increased significantly (p<0.05) during storage
indicating hydrolysis and degradation of protein. Similarly, showing high hydrolytic rancidity, the peroxide value (PV), free
fatty acid (FFA) and thiobarbituric acid (TBA) number significantly increased throughout the storage period (p<0.05). The
sensory scores showed significant differences (p<0.05) during the storage period. Shidal retained good quality up to 60 days at
room temperature whereas 90 days at refrigerated temperature showing significantly high sensory scores in the treatment.

Full text

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Extension of shelflife of the fermented fish product, shidal by packaging
in glass bottle and low temperature storage
P. MAHANTA AND A. U. MUZADDADI
College of Fisheries, Central Agricultural University, Lembucherra – 799 210, Tripura, India
e-mail: drarmaan@yahoo.com
ABSTRACT
Shidal is a very popular, salt free, semi-fermented traditional fish product of north-east India prepared using minor carps
(Puntius spp.) in specially designed earthen pots (mutka). The prime quality of shidal is given by its characteristic aroma and
flavor which is lost very fast, once it is taken out of mutka. A study was conducted to preserve the quality of shidal outside
mutka, by packing in glass bottles and storing under refrigerated temperature. Microbiological, biochemical and sensory changes
during storage period of 120 days were analysed at 15 days interval. The total plate count (TPC) did not change significantly
(p>0.05) and remained near 7 log CFU g
-1
during storage. The total fungal count (TFC) was negligible., The pH of shidal was
initially acidic (4.42 ± 0.25) which increased significantly (p<0.05) towards the end of storage period. The non-protein nitrogen
(NPN), free α-amino nitrogen (AAN) and total volatile base nitrogen (TVB-N) increased significantly (p<0.05) during storage
indicating hydrolysis and degradation of protein. Similarly, showing high hydrolytic rancidity, the peroxide value (PV), free
fatty acid (FFA) and thiobarbituric acid (TBA) number significantly increased throughout the storage period (p<0.05). The
sensory scores showed significant differences (p<0.05) during the storage period. Shidal retained good quality up to 60 days at
room temperature whereas 90 days at refrigerated temperature showing significantly high sensory scores in the treatment.
Keywords: Biochemical composition, Fermented fish, Puntius spp., Shidal, Storage characteristics
Introduction
Shidal is a salt free, semi- fermented and very popular
fish product of north-east India. With high rancid odour
and prepared from small carps mainly Puntius spp. shidal
is an unavoidable fish-based product as far as food security
of the people of north-east is concerned. There are several
similar products in different north-eastern states such as
seedal and hidal in Assam, sepaa and shidal in Tripura,
Nagaland and Arunachal Pradesh and ngari in Manipur
(Muzaddadi, 2002; 2003a).The fish is semi dried and almost
anaerobically fermented in earthen pots. The fermentation
process takes around four to six months in anaerobic
condition till the product gains a characteristic odour,
texture and appearance. Neither food additives/
preservatives nor starter culture is added during the
processing steps (Muzaddadi and Basu, 2003b). This
product is also popular in Bangladesh which is known as
chepa shutki (Nayeem et al., 2010). Sarojnalini and
Viswanath (1998) reported about almost similar type of
fermented products in Manipur viz., hentak and ngari,
which are prepared from sun-dried freshwater fish.
The traditional products are generally contaminated
with dirt, filth, sand and dust during retail marketing.
Bamboo basket, jute bags and earthen pots are used for
storing the fermented fish. The materials are usually used
under poor sanitation and hygiene and these products often
Indian J. Fish., 60(2) : 135-143, 2013
9
turn brown to dark brown in colour and are heavily infested
with insects. Shidal has a very short storage life once it is
taken out from the mutka. Therefore, mutkas are used as
primary packaging and transporting vessels. Since glass is
inert material having highest impermeability to gas and
volatile substances, glass bottles might act as effective
packaging material which is expected to retain the volatile
flovour components of shidal. Though shidal is very
common in every household of north-eastern parts of India,
scientific information regarding storage are very scanty.
Moreover, shidal has not been explored scientifically in
India. The present study is expected to provide a
comprehensive information on low cost packaging method
for retailing and house hold storage of shidal. The study
aimed to analyse the preservative action of low temperature,
under packaged condition.
Materials and methods
The study was carried out in the fish processing
technology laboratory, College of Fisheries, Central
Agricultural University, Lembucherra, Tripura west.
Required amount of first grade commercial shidal prepared
using four species of Puntius (P. chola, P. sarana,
P. sophore and P. ticto) were collected aseptically in sterile
polyethylene bags immediately after opening the mutka
from the production centres in Agartala. For storage study,
2 kg shidal was packed in glass bottles (3l capacity,

136
19.5 cm X 13.5 cm height X, 10 cm outer mouth dia and
0.5 cm glass thickness, with cork and polythene lined
stainless steel screw caps) and then stored at 4 ºC (T) and
also at ambient temperature separately which served as
control (C) (Table 1).
Table 1. Sampling schedule of shidal along with ambient
temperature recorded
Sampling Date Ambient Temperature (
0
C)
1 20 August 2010 30-34
2 5 September 2010 28-33
3 20 September 2010 28-33
4 5 October 2010 24-28
5 20 October 2010 24-28
6 5 November 2010 20-28
7 20 November 2010 20-28
8 5 December 2010 18-25
9 20 December 2011 18-25
The sampling was done aseptically in sterile
petridishes, from each glass bottles for different
microbiological, biochemical and sensory analysis. The
container was closed immediately after sampling and the
sampling was continued at 15 days’ interval up to 120 days
of storage.
Total plate count (TPC) and total fungal count (TFC)
were done by spread plate technique (APHA, 1995). For
this, 10 g of shidal sample was introduced aseptically in a
sterile stomacher bag (Seward stomach BA6141CPG
standard bags) and macerated for 2 min with 90 ml of sterile
diluent (0.85% NaCl) using a stomacher (Seward stomacher
400 circulator, England). Serial dilutions were made and
plated onto Soybean Casein Digest Agar (SCDA,
HIMEDIA) for TPC and onto Rose Bengal Chloramphenical
agar (RBCPA, HIMEDIA) plates for TFC.
The samples were analysed in triplicate for moisture,
ash, pH and free fatty acids (FFA) following AOAC (2000),
lipid content by the Soxlet method (AOAC, 2000), protein
and non-protein nitrogen (NPN) by the Kjeldahl method
(AOAC, 2000); total volatile base nitrogen (TVB-N)
content according to the Conway’s micro-diffusion method
(Conway, 1947); the thiobarbituric acid (TBA) value
following Tarladgis et al. (1960); peroxide value (PV) as
per Jacob (1958); and the free
α
-amino nitrogen (AAN)
by the method of Pope et al. (1939).
Sensory studies of shidal were carried out by an expert
panel of 10 expert judges by 5-points Hedonic scale
(Table 2). The overall acceptability was calculated by taking
arithmetic average from score-sheet.
Statistical analysis
Statistical analysis was done by performing one way
ANOVA (Post Hoc, Duncan) and student’s t-test to compare
the means using SPSS 15.0 (2005) at 5% confidence level.
All bacteriological counts were converted to log10
CFU g
-1
for statistical analysis.
Results and discussion
Total plate count (TPC) was analysed during the study
period to see the effect of temperature on the survival of
Table 2. Quality scores for the sensory evaluation of shidal
Quality attributes Characteristics Sensory scores Quality
Appearance Bright, moist and shining surface 5 Excellent
Slight dullness with shining surface 4 Good
Dull with soft surface 3 Fair
Definite dullness with soft surface 2 Average
Dry fish like, loss of weight, fungal growth 1 Poor
Colour Dark brownish or slight yellowish colour 5 Excellent
Gray yellowish or black colour 4 Good
Pale brown or gray colour 3 Fair
Colour become fade and off-white 2 Average
Whitish colour 1 Poor
Odour Strong characteristic Shidal odour 5 Excellent
Light characteristic Shidal odour 4 Good
Slight Shidal odour with no sour odour 3 Fair
Faint sour odour 2 Average
Strong sour odour, rancid, ammonia smell 1 Poor
Texture Firm elastic muscle, sticky surface and muscle not 5 Excellent
detached from the backbone
Soft abdomen and fairly firm muscle 4 Good
Muscle with no elasticity and firmness 3 Fair
Limp and flaccid muscle 2 Average
Melting abdomen, muscle easily bruised and broken 1 Poor
P. Mahanta and A. U. Muzaddadi

137
bacteria under refrigerated condition in comparison with
ambient temperature storage of 120 days and expressed as
log CFU g
-1
. The TPC of control (C) showed significant
differences (p<0.05) with that of the treatment (T)
(Fig. 1).. Initial decrease in TPC was recorded in T, which
may be due to the cold shock to the mesophilic bacteria at
lower temperature and the psychrotrophic bacteria took time
under chilled temperature for adapting themselves to cold
environment and the growth was observed during further
storage period. Similar observations were reported in
fermented cassava fish wherein the mesophilic bacterial
count decreased from 6.25 to 4.94 log CFU g
-1
during initial
fermentation (Anihouvi et al., 2007). It was observed that
the counts in C and T were almost same during initial
15 days of storage and subsequently the count of T became
higher than that of C. During the end of the storage period
the count in T was significantly (p<0.05) different from
that of C. The reason for this may be attributed to the
overgrowth of bacteria in C at ambient temperature, which
resulted in reaching the lag phase of growth during the
end of the storage period. However, low temperature
preservation retarded the growth of psychrotrophic and
mesophilic bacteria which in turn, prolonged the log phase
and hence the growth of bacteria continued till the end of
the storage period. Sarojnalini and Suchitra (2009) reported
a similar increasing trend of Gram positive bacteria in
fermented Setipinna sp. of Manipur. The findings of Thapa
and Tamang (2004) in ngari, hentak and tungtap of
north-east India also agree with the present findings.
Total fungal counts (TFC) recorded were <2500 CFU g
-1
in most of the samples, and in many samples TFC was
undetectable. Similar trend was observed by Anihouvi et
al. (2007) in fermented cassava fish.
Proximate composition (Table 3) clearly showed that
shidal is a highly nutritious food item. The pH showed
significant differences (p<0.05) during the storage period
(Fig. 2). The initial decrease in pH indicates fermentation
process and formation of organic acids. The decrease in
pH may also be due to the high buffering capacity of the
fish flesh (Dakwa et al., 2005). Nevertheless, fermentation
Shelflife of the fermented fish product, shidal
Table 3. Desirable microbial and biochemical quality parameters of shidal
Parameters Values (mean + SD)
Total plate count (log CFU g
-1
) 7.1 ± 0.12
Total fungal count (log CFU g
-1
) ETFC <2500
pH 4.42 ± 0.43
Moisture (w/w %) 34.02 ± 1.55
Ash (w/w %) 13.80 ± 0.32
Protein (w/w %) 31.06 ± 0.05
Fat (w/w %) 18.87 ± 0.42
Acid-insoluble ash (w/w %) 0.53 ± 0.02
Non-protein nitrogen (w/w %) 5.49 ± 0.24
Free alpha amino acid (w/w mg %) 6.67 ± 4.67
Total volatile base nitrogen (mg %) 223.67 ± 3.28
Peroxide value (milliequivallent peroxide oxygen per 1000 g) 17.03 ± 0.32
Free fatty acid (% as oleic acid) 20.62 ± 0.17
Thiobarbituric acid number (mg malonaldehyde per 1000 g) 0.51 ± 0.01
Fig. 1. Changes in TPC (log CFU g
-1
) of shidal during storage,
values = mean ± SE (error bars), n = 3
Fig. 2. Changes in pH of shidal during storage,
values = mean ± SE (error bars), n = 3

138
could not continue for longer period due to the absence of
fermenting bacteria in the later stages due to aerobic storage
condition. Thus, in the later period of storage, fermentation
stopped and protein was further degraded to form some
volatile bases which led to increased pH. It may be
attributed to putrefaction leading to formation of basic
nitrogenous compounds (Kilinc et al., 2006). Low
temperature influenced the pH value which may be due to
restriction in growth of fermenting bacteria. These findings
agree with that of Sarojnalini and Suchitra (2009) in
fermented fish of Manipur.
There were significant differences in moisture and ash
(p<0.05) (Fig. 3 and 4). The moisture content decreased
gradually and the reason may be the initial low pH which
electrically neutralised the proteins which might have
caused decrease in water holding capacity of fish meat
(Akahane and Shimizu, 1989). Itou et al. (2006) also
reported a decreasing trend in moisture in narezushi
(a Japanese fermented mackerel product) during processing.
The ash content was too high which indicates sand
adulteration as well as unhygienic conditions during
preparation in traditional production centers. Sarojnalini
and Suchitra (2009) also reported high content of ash in
fermented Setipinna sp. of Manipur. Majumdar et al. (2006)
reported similar trends in ash content of lona ilish (salted
and fermented hilsa (Tenualosa ilisha) from north-east
India) during fermentation.
Being the major component of shidal, protein content
differed significantly (p<0.05) during storage (Fig. 5) and
gradually decreased at a constant rate during entire storage
period. It may be due to hydrolysis of protein by intrinsic
and microbial enzymatic action (Majumdar et al., 2006).
The initial decreases in protein percentage may be due to
the increment in total weight by the increased moisture
content. Nayeem et al. (2010) also reported similar decrease
in protein content of chepa shutki collected from producer,
whole-seller and retailer, with increasing storage period.
Some volatile nitrogenous compounds might escape to the
atmosphere due to which a reduction in total nitrogen
content resulted and thus reduction in protein content.
Similar observation was also reported by Taira et al. (2007)
during fermentation of fish sauce.
P. Mahanta and A. U. Muzaddadi
Fig. 3. Changes in moisture content of shidal during storage
values = mean ± SE (error bars), n = 3
Fig. 4. Changes in ash content of shidal during storage
values = mean ± SE (error bars), n =3
Fig. 5. Changes in protein content of shidal during storage
values = mean ± SE (error bars), n = 3
NPN, AAN and TVB-N showed significant
differences (p<0.05) with increasing storage period. The
significant increase in all these products may be the result
of degradation of protein during storage (Fig. 6, 7 and 8).
The TVB-N content was observed high (>200 mg%) in the
treatment and differed significantly from control (p<0.05).
The findings of Karthikeyan et al. (2007) and Majumdar
and Basu (2010) agree with the present findings. TMA-N
and TVB-N are products of bacterial spoilage and the
content is often used as an index to assess the keeping
quality and shelflife of seafood products (Vareltzis et al.,
1997). Karacam et al. (2002) reported similar increasing
trend in TVB-N showing no temperature effect on brined

139
Shelflife of the fermented fish product, shidal
Fig. 9. Changes in fat content of shidal during storage
values = mean ± SE (error bars), n = 3
Fig. 10. Changes in peroxide value (PV) of shidal during storage
values = mean ± SE (error bars), n = 3
Fig. 11.Changes FFA content of shidal during storage
values = mean ± SE (error bars), n = 3
Fig 12. Changes in TBA number of shidal during storage
values = mean ± SE (error bars), n = 3
Fig. 6. Changes in NPN of shidal
during storage, values = mean ±S E
(error bars), n = 3
Fig. 7. Changes in AAN of shidal
during storage, values = mean ± SE
(error bars), n = 3
Fig. 8. Changes in TVBN content of
shidal during storage,
values =
mean ±S E (error bars), n = 3
anchovies. Ndaw et al. (2008) also reported increased
TVB-N values in fermented Moroccan sardines (Sardinella
pilchardus). TVB-N values recorded in the present study
clearly indicate deterioration of proteins. The increase in
AAN throughout the period may be because of the
combined effect of autolysis and microbial degradation of
the fish muscle (Voskresensky, 1965; Ijong and Ohta, 1996).
This may be implicated in the activities of enzymes which
originated from fish gut, muscle and bacteria (Majumdar
et al., 2006). The contents of NPN and TVB-N of Shidal
were indicative of high degree of fermentation.
The lipid content and different lipid degraded products
like PV, FFA and TBA showed significant differences
(p<0.05) during the storage period (Fig. 9 - 12). The fat
content gradually decreased throughout the period, may

140
be due to oxidative and hydrolytic rancidity of fat during
storage. Considerable decomposition of triglyceride and
phospholipids in the lipid may occur, accompanied by the
production and accumulation of large amounts of free fatty
acids throughout processing (Kilinc et al., 2006). The high
amount of fat may be due to additional fish oil which is
normally used for smearing mutka and the raw material,
and also Puntius spp. contain high amount of fat and they
are caught during monsoon period (Sarojnalini and
Vishwanath, 1994). The decreasing trend in fat content was
also observed by Karthikeyan et al. (2007) in smoked
P. Mahanta and A. U. Muzaddadi
Colisa fasciata of Manipur, by Rahman et al. (1999) in
salted Hilsa sp. and by Nayeem et al. (2010) in chepa shutki
with increasing storage time. There are formal chemical
definitions of oxidation, involving electron transfer and free
radical reactions but, in the context of fish technology, it
can be considered as the chemical reaction in which oxygen
combines with a compound (Kilinc et al., 2006). Among
the different lipid degraded products, the primary oxidation
indicator PV which is the index to assess the lipid oxidation
of shidal significantly differed (p<0.05) in the treatment
and the control (Fig. 10). The gradual increase in PV values
Fig. 13. Changes in sensory scores for different sensory parameters (appearance, colour, texture, odour and overall acceptability) of
shidal during 120 days of storage, values = mean ± SE (error bars), n = 10

141
Shelflife of the fermented fish product, shidal
may be due to more amount of initial fat oxidation and it
also indicates the formation of peroxide or hydroperoxide
(Vernam and Sutherland, 1995).
The FFA content increased throughout the storage
period (Fig. 11). FFA showed less values in refrigerated
samples indicating that low temperature storage could
reduce lipid hydrolysis. These findings agree with the
findings of Srikar et al. (1993) who reported lower PV and
lower content of FFA in salted mackerel and pink perch
when stored at 2.5
o
C after 35 days of storage. The high
TBA values represent the degree of rancidity in the products
and the values above 3-4 indicate quality loss (Karacam
and Boran, 1996). It was observed that in the present study,
the values are in the acceptable range.
Primary and secondary lipid oxidation products are
the biological amino compounds, protein, peptides, free
amino acids and phospholipids; they react to produce
interaction compounds and this make the colour of the
product brown, causes a change in flavour and loss in
aromatic nutrient elements (Aubourg, 1998).
The sensory score for all sensory characters
significantly differed (p<0.05). The scores for appearance,
colour, texture, odour and overall acceptability (Fig. 13)
of shidal significantly differed (p<0.05) and gradually
decreased at a higher rate.
It was observed that the low temperature storage of
shidal had positive effects on extending shelflife of shidal.
Low temperature stored shidal showed less fluctuations in
the scores of all the sensory parameters which denotes a
stable quality of shidal during the storage period (Fig. 13).
The product remained acceptable up to 90 days of storage
with an overall acceptability score of >3.0. Shidal after
90 days storage under refrigerated condition is shown in
Fig. 14. This agrees with the findings of Akande et al. (1991)
in spiced minced fish cake and Koral et al. (2010) in hot
smoked Atlantic bonito (Sarda sarda) packed in aluminium
foil during storage. Ozden and Erkan (2006) also reported
that sensory evaluation has an important role in determining
the quality of fish and seafood. After analysing and
observing all the parameters it was inferred that shidal
may be kept at low temperature up to 90 days without losing
its characteristic odour and biochemical quality. It became
unacceptable after 120 days of storage under both
conditions (Fig. 15).
Fig. 14. Good quality Shidal after 90 days of storage under
refrigerated condition
Fig. 15. Shidal after 120 days of storage
Though shidal is one of the important fermented
products available in north-eastern part of India, it does
not have proper packaging and preservation methods and
it loses its typical flavour within a short period after taking
out from the mutka. The present study expected to provide
basic scientific information to develop a suitable packaging
and preservation technology for shidal. Limited research
work has been done on all aspects of fermented fish
products of north-east India. and there is a need for future
research especially in developing better packaging methods
utilising polythene, polyethylene terephthalate (PET) or
modified atmospheric packaging (MAP).
Acknlowledgements
Authors are thankful to the Vice Chancellor, Central
Agricultural University and Dean, College of Fisheries,
Agartala for providing infrastructural and financial support
to carry out the research.
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Date of Receipt : 03.09.2011
Date of Acceptance : 26.12.2012